The Dynamics of DNA Damage Repair and Transcription

Shanbhag, Niraj M.; Greenberg, Roger A.

doi:10.1007/978-1-62703-526-2_16

Cited by 14 publications

(15 citation statements)

References 21 publications

(24 reference statements)

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“…Nascent transcription is visualized by the accumulation of fluorescent MS2-binding proteins at the transcription site, upon binding to nascent MS2 stem-loop structures present at the reporter gene RNA (Shanbhag et al, 2010). Expression of the FokI nuclease domain fused to the mCherry-LacI creates DSBs at the lac operator array (Shanbhag and Greenberg, 2013). Of note, this approach leads to persistent and extensive DSB induction and the time of damage induction is dependent on the expression of mCherry-LacI-FokI.…”

Section: Methods To Induce Annotated Dna Breaks At Transgenic Loci Inmentioning

confidence: 99%

Studying DNA Double-Strand Break Repair: An Ever-Growing Toolbox

Vítor

Huertas

Legube

et al. 2020

Front. Mol. Biosci.

118

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To ward off against the catastrophic consequences of persistent DNA double-strand breaks (DSBs), eukaryotic cells have developed a set of complex signaling networks that detect these DNA lesions, orchestrate cell cycle checkpoints and ultimately lead to their repair. Collectively, these signaling networks comprise the DNA damage response (DDR). The current knowledge of the molecular determinants and mechanistic details of the DDR owes greatly to the continuous development of groundbreaking experimental tools that couple the controlled induction of DSBs at distinct genomic positions with assays and reporters to investigate DNA repair pathways, their impact on other DNAtemplated processes and the specific contribution of the chromatin environment. In this review, we present these tools, discuss their pros and cons and illustrate their contribution to our current understanding of the DDR.

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Section: Methods To Induce Annotated Dna Breaks At Transgenic Loci Inmentioning

confidence: 99%

Studying DNA Double-Strand Break Repair: An Ever-Growing Toolbox

Vítor

Huertas

Legube

et al. 2020

Front. Mol. Biosci.

118

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“…As chromatin is actively reorganized around sites of damage to facilitate repair, we next determined whether FKBP25 is actively recruited to DNA double-strand breaks. We evaluated co-localization of an mCherry-LacI-FokI fusion and FKBP25-GFP at an induced DNA break near an integrated lacO array ( Shanbhag and Greenberg 2013 ) and did not observe colocalization of FKBP25 at DSBs (data not shown). We next tested whether FKBP25 might visit DNA lesions transiently.…”

Section: Resultsmentioning

confidence: 99%

FKBP25 participates in DNA double-strand break repair

Dilworth

Gong

Miller

et al. 2020

Biochem. Cell Biol.

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FK506-binding proteins (FKBPs) alter the conformation of proteins via cis–trans isomerization of prolyl-peptide bonds. While this activity can be demonstrated in vitro, the intractability of detecting prolyl isomerization events in cells has limited our understanding of the biological processes regulated by FKBPs. Here we report that FKBP25 is an active participant in the repair of DNA double-strand breaks (DSBs). FKBP25 influences DSB repair pathway choice by promoting homologous recombination (HR) and suppressing single-strand annealing (SSA). Consistent with this observation, cells depleted of FKBP25 form fewer Rad51 repair foci in response to etoposide and ionizing radiation, and they are reliant on the SSA repair factor Rad52 for viability. We find that FKBP25’s catalytic activity is required for promoting DNA repair, which is the first description of a biological function for this enzyme activity. Consistent with the importance of the FKBP catalytic site in HR, rapamycin treatment also impairs homologous recombination, and this effect is at least in part independent of mTor. Taken together these results identify FKBP25 as a component of the DNA DSB repair pathway.

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“…Human HEK293T (fetal), U2OS (female), and HCT116 (male) cell lines were originally purchased from the American Type Culture Collection (ATCC, Manassas, VA). U2OS-235 mCherry-LacI-FokI cell lines (female) were kindly provided by Dr. Roger Greenberg and have been described previously ( Shanbhag and Greenberg, 2013 ). U2OS-DR-GFP cell lines were kindly provided by Dr. Jeremy Stark and have been described previously ( Pierce et al, 1999 ).…”

Section: Methodsmentioning

confidence: 99%

SIRT2 promotes BRCA1-BARD1 heterodimerization through deacetylation

Kapoor-Vazirani

Zhang

et al. 2021

Cell Reports

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SUMMARY The breast cancer type I susceptibility protein (BRCA1) and BRCA1-associated RING domain protein I (BARD1) heterodimer promote genome integrity through pleiotropic functions, including DNA double-strand break (DSB) repair by homologous recombination (HR). BRCA1-BARD1 heterodimerization is required for their mutual stability, HR function, and role in tumor suppression; however, the upstream signaling events governing BRCA1-BARD1 heterodimerization are unclear. Here, we show that SIRT2, a sirtuin deacetylase and breast tumor suppressor, promotes BRCA1-BARD1 heterodimerization through deacetylation. SIRT2 complexes with BRCA1-BARD1 and deacetylates conserved lysines in the BARD1 RING domain, interfacing BRCA1, which promotes BRCA1-BARD1 heterodimerization and consequently BRCA1-BARD1 stability, nuclear retention, and localization to DNA damage sites, thus contributing to efficient HR. Our findings define a mechanism for regulation of BRCA1-BARD1 heterodimerization through SIRT2 deacetylation, elucidating a critical upstream signaling event directing BRCA1-BARD1 heterodimerization, which facilitates HR and tumor suppression, and delineating a role for SIRT2 in directing DSB repair by HR.

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The Dynamics of DNA Damage Repair and Transcription

Cited by 14 publications

References 21 publications

Studying DNA Double-Strand Break Repair: An Ever-Growing Toolbox

Studying DNA Double-Strand Break Repair: An Ever-Growing Toolbox

FKBP25 participates in DNA double-strand break repair

SIRT2 promotes BRCA1-BARD1 heterodimerization through deacetylation

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